primary antibody rabbit anti human ki67  (Cell Signaling Technology Inc)

Journal: Oncotarget

Article Title: Expression of receptor activator of NFkB (RANK) drives stemness and resistance to therapy in ER+HER2- breast cancer

doi: 10.18632/oncotarget.27576

Figure Lengend Snippet: ( A ) BLI analysis of MCF-7 and MCF-7 OE xenografts in NSG mice ( n = 5–8/group). ( B ) IHC analysis of Ki67. ( C ) Microvessel density (MVD) was quantified after VWF immunohistochemistry. ( D ) Predicted proliferation rate in TCGA cohort according to median RANK expression in each sub-group. ( E ) Ex vivo BLI of visceral organs and quantification of lung tumor burden. ( F ) BLI analysis of mice inoculated in the tail vein with MCF-7 or MCF-7 OE cells ( n = 5/group). ( G ) Flow cytometry analysis of positive GFP cells in whole blood collected at sacrifice. ( H , I ) Tumor volume (Tvol = 1/2 (length × width 2 )) measured at necropsy of T47D or T47D OE adherent cell-derived xenografts ( n = 3/group) (H) or T47D or T47D OE tumorspheres-derived xenografts ( n = 5/group) (I). FiJi was used to obtain the best contrast for western blot band visualization, and background was removed for band densitometry analysis. Data is presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Specific antibodies included rabbit anti-human Ki67 primary antibody (1:100, MIB-1, Dako), rabbit anti-human ERα (RTU, EP-1, Dako) and polyclonal rabbit anti-human von Willebrand Factor (VWF) primary antibody (1:200, #A0082, Dako), and EnVisionTM Detection System, rabbit/mouse (#411083, Dako).

Techniques: Immunohistochemistry, Expressing, Ex Vivo, Flow Cytometry, Derivative Assay, Western Blot

Journal: Oncotarget

Article Title: Expression of receptor activator of NFkB (RANK) drives stemness and resistance to therapy in ER+HER2- breast cancer

doi: 10.18632/oncotarget.27576

Figure Lengend Snippet: ( A ) BLI analysis of MCF-7 and MCF-7 OE xenografts in NSG mice ( n = 3–4/group). ( B ) Tumors photographed at necropsy. ( C ) Tumor volume (Tvol = 1/2 (length × width 2 )) measured at necropsy. ( D ) Tumor weight at necropsy. ( E ) IHC analysis of Ki67. ( F ) Cell viability was measured 6 weeks after exposure to RANKL ( n = 3). ( G ) Western blot of RANK downstream targets and cell cycle control proteins with β-Actin as loading control. ( H ) RT-qPCR of RANK ( n = 3). ( I ) Flow cytometry of RANK. ( J , K ) Western blot of indicated proteins with β-Actin as loading control. ( L ) Interest gene expression in the ER+HER2- TCGA cohort ( n = 587) according to RANK expression. ( M ) Western blot of indicated proteins with β-Actin as loading control. ( N ) Representative images of IHC of ER. ( O ) Cell viability was measured 7 days after exposure to the indicated drugs, with medium replacement every 48 h. ( n = 3). FiJi was used to obtain the best contrast for western blot band visualization, and background was removed for band densitometry anslysis. Data is presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Specific antibodies included rabbit anti-human Ki67 primary antibody (1:100, MIB-1, Dako), rabbit anti-human ERα (RTU, EP-1, Dako) and polyclonal rabbit anti-human von Willebrand Factor (VWF) primary antibody (1:200, #A0082, Dako), and EnVisionTM Detection System, rabbit/mouse (#411083, Dako).

Techniques: Western Blot, Quantitative RT-PCR, Flow Cytometry, Expressing

Journal: Investigative Ophthalmology & Visual Science

Article Title: Effect of Methotrexate on an In Vitro Patient-Derived Model of Proliferative Vitreoretinopathy

doi: 10.1167/iovs.16-20912

Figure Lengend Snippet: Effect of MTX on proliferation of C-PVR cells. Immunofluorescence images of C-PVR cells from after 24, 48, and 72 hours without (A, E, I) and with 100 (B, F, J), 200 (C, G, K), and 400 mM (D, H, L) MTX. Quantification of proliferation by Ki67 immunofluorescence (red) at 24, 48, and 72 hours (M–O, respectively). Error bars represent SEM. Scale bar denotes 100 μm.

Article Snippet: Primary antibody rabbit anti-human Ki67 antibody (1:100; Novus Biologicals) and cell nuclei were labeled with Hoechst 33342 diluted in PBS 1:200 (ImmunoChemistry Technologies).

Techniques: Immunofluorescence